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Pleiotropic corepressors Sin3 and Ssn6 interact with repressor Opi1 and negatively regulate transcription of genes required for phospholipid biosynthesis in the yeast Saccharomyces cerevisiae

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Abstract

Repressor protein Opi1 is required to negatively regulate yeast structural genes of phospholipid biosynthesis in the presence of precursor molecules inositol and choline (IC). Opi1 interacts with the paired amphipathic helix 1 (PAH1) of pleiotropic corepressor Sin3, leading to recruitment of histone deacetylases (HDACs). Mutational analysis of the Opi1–Sin3 interaction domain (OSID) revealed that hydrophobic OSID residues L56, V59 and V67 of Opi1 are indispensable for gene repression. Our results also suggested that repression is not executed entirely via Sin3. Indeed, we could show that OSID contacts a second pleiotropic corepressor, Ssn6 (=Cyc8), which together with Tup1 is also able to recruit HDACs. Interestingly, mutations sin3 and ssn6 turned out as synthetically lethal. Our analysis further revealed that OSID not only binds to PAH1 but also interacts with tetratricopeptide repeats (TPR) of Ssn6. This interaction could no longer be observed with Opi1 OSID variants. To trigger gene repression, Opi1 must also interact with activator Ino2, using its activator interaction domain (AID). AID contains a hydrophobic structural motif reminiscent of a leucine zipper. Our mutational analysis of selected positions indeed confirmed that residues L333, L340, V343, V350, L354 and V361 are necessary for repression of Opi1 target genes.

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Acknowledgments

This work has been supported by the Deutsche Forschungsgemeinschaft (DFG). We thank Felix Kliewe, Gudrun Ebel and Karola Hahn for valuable support.

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  1. Institut für Genetik und Funktionelle Genomforschung, Jahnstrasse 15a, 17487, Greifswald, Germany

    Yvonne Jäschke, Juliane Schwarz, Diana Clausnitzer, Carina Müller & Hans-Joachim Schüller

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  1. Yvonne Jäschke
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  2. Juliane Schwarz
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  3. Diana Clausnitzer
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Correspondence to Hans-Joachim Schüller.

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Communicated by M. Collart.

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Jäschke, Y., Schwarz, J., Clausnitzer, D. et al. Pleiotropic corepressors Sin3 and Ssn6 interact with repressor Opi1 and negatively regulate transcription of genes required for phospholipid biosynthesis in the yeast Saccharomyces cerevisiae . Mol Genet Genomics 285, 91–100 (2011). https://doi.org/10.1007/s00438-010-0589-5

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